Enhanced Thermal Stability/Activity of Geobacillus jurassicus Esterase by Rational Design and Application in the Synthesis of Cinnamyl Acetate.

IF 3.1 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Junze Li, Runfei Song, Lin Lin, Tao Li, Yan Yan, Wei Wei, Dongzhi Wei
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引用次数: 0

Abstract

Geobacillus sp. represents an important source of thermophilic esterases, yet studies on the rational design and industrial application of these enzymes remain limited. In our previous research, we identified the esterase Gju768 from Geobacillus jurassicus DSMZ 15726. In the present study, we employed a novel computer-aided rational design approach, ACDP (AutoDock, Consurf, Discovery Studio, PoPMuSiC), to enhance the enzyme's thermal stability. Through molecular docking and conservation analysis, three hotspots were identified. Virtual saturation mutagenesis was subsequently performed, yielding two selected mutations, Q78I and Q78L, from the resulting library. Notably, mutants Q78I and Q78L exhibited significant improvements in thermal stability and enzyme activity compared to the wild type (WT). Compared to WT, mutants Q78I and Q78L exhibited a 65.27% and 38.38% increase in half-life at 65 °C, along with a 14.48% and 1.60% improvement in specific activity at their respective optimal temperatures. Furthermore, under optimized conditions for cinnamyl acetate production, mutant Q78I demonstrated a yield of 68%, compared to only 31% for WT. This study underscored the potential of protein engineering strategies to enhance enzyme performance in industrial applications, particularly for the synthesis of value-added compounds such as cinnamyl acetate.

合理设计提高侏罗纪地杆菌酯酶热稳定性/活性及其在肉桂酯合成中的应用
Geobacillus sp.是一种重要的嗜热酯酶来源,但对这些酶的合理设计和工业应用的研究仍然有限。在之前的研究中,我们从侏罗纪地杆菌DSMZ 15726中鉴定出酯酶Gju768。在本研究中,我们采用了一种新的计算机辅助理性设计方法ACDP (AutoDock, Consurf, Discovery Studio, PoPMuSiC)来提高酶的热稳定性。通过分子对接和保守分析,确定了三个热点。随后进行虚拟饱和诱变,从生成的文库中产生两个选定的突变,Q78I和Q78L。值得注意的是,与野生型(WT)相比,突变体Q78I和Q78L在热稳定性和酶活性方面表现出显著改善。与WT相比,突变体Q78I和Q78L在65°C时的半衰期分别提高了65.27%和38.38%,比活性分别提高了14.48%和1.60%。此外,在优化的肉桂乙酸酯生产条件下,突变体Q78I的产量为68%,而WT的产量仅为31%。该研究强调了蛋白质工程策略在工业应用中提高酶性能的潜力,特别是在合成肉桂乙酸酯等增值化合物方面。
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来源期刊
Applied Biochemistry and Biotechnology
Applied Biochemistry and Biotechnology 工程技术-生化与分子生物学
CiteScore
5.70
自引率
6.70%
发文量
460
审稿时长
5.3 months
期刊介绍: This journal is devoted to publishing the highest quality innovative papers in the fields of biochemistry and biotechnology. The typical focus of the journal is to report applications of novel scientific and technological breakthroughs, as well as technological subjects that are still in the proof-of-concept stage. Applied Biochemistry and Biotechnology provides a forum for case studies and practical concepts of biotechnology, utilization, including controls, statistical data analysis, problem descriptions unique to a particular application, and bioprocess economic analyses. The journal publishes reviews deemed of interest to readers, as well as book reviews, meeting and symposia notices, and news items relating to biotechnology in both the industrial and academic communities. In addition, Applied Biochemistry and Biotechnology often publishes lists of patents and publications of special interest to readers.
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